AW: [Histonet] Movat Pentachrome - Need to enhance Alcian Blue
Hi Krista, Maybe the bad stainability is more due to oxidation by air of the old block-surface than due to fixation. A deeper slide for comparison would show. I don't know what the alkaline alcohol is good for ? Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Krista Sider Gesendet: Montag, 27. Mai 2013 23:28 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Movat Pentachrome - Need to enhance Alcian Blue Hi All, I had wonderful help from this site for my Woodstain Scarlet-Acid Fuchsin binding collagen problem. Now that this is fixed I have realized that the Alcian Blue is not staining my excessively fixed samples (Human Aorta Aortic Valve, formalin fixed 2-4 years, embedded in paraffin and stored at RT for ~6 years). I have some samples fixed for only 2-6 months and these are fine but the samples fixed for 2-4 years do not uptake the Alcian Blue. I have taken it up to 1 hr in the 1% Alcian Blue solution followed by 10 min in 56*C Alkaline Alcohol solution. I do a 1hr Bouin's 50*C to start (I have also tried Bouin's at 60*C). Any ideas on how I can get the Alcian Blue to stain these excessively fixed samples or possibly a substitute? I need to see the proteoglycans/GAGs. Thank you Krista ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: AW: [Histonet] Movat Pentachrome - Need to enhance Alcian Blue
Hi Gudrun How deep would you consider the oxidation to go? I have tried samples from around 200um into the block. The Alkaline Alcohol converts the alcian blue to monastral fast blue, which is insoluble - this is what i have been told. So locks it in I guess. Doesn't really matter as I don't get alcian blue in the sample to lock in. Thanks Krista On May 28, 2013 10:09 AM, Gudrun Lang gu.l...@gmx.at wrote: Hi Krista, Maybe the bad stainability is more due to oxidation by air of the old block-surface than due to fixation. A deeper slide for comparison would show. I don't know what the alkaline alcohol is good for ? Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Krista Sider Gesendet: Montag, 27. Mai 2013 23:28 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Movat Pentachrome - Need to enhance Alcian Blue Hi All, I had wonderful help from this site for my Woodstain Scarlet-Acid Fuchsin binding collagen problem. Now that this is fixed I have realized that the Alcian Blue is not staining my excessively fixed samples (Human Aorta Aortic Valve, formalin fixed 2-4 years, embedded in paraffin and stored at RT for ~6 years). I have some samples fixed for only 2-6 months and these are fine but the samples fixed for 2-4 years do not uptake the Alcian Blue. I have taken it up to 1 hr in the 1% Alcian Blue solution followed by 10 min in 56*C Alkaline Alcohol solution. I do a 1hr Bouin's 50*C to start (I have also tried Bouin's at 60*C). Any ideas on how I can get the Alcian Blue to stain these excessively fixed samples or possibly a substitute? I need to see the proteoglycans/GAGs. Thank you Krista ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Movat Pentachrome - Need to enhance Alcian Blue
Hi All, I had wonderful help from this site for my Woodstain Scarlet-Acid Fuchsin binding collagen problem. Now that this is fixed I have realized that the Alcian Blue is not staining my excessively fixed samples (Human Aorta Aortic Valve, formalin fixed 2-4 years, embedded in paraffin and stored at RT for ~6 years). I have some samples fixed for only 2-6 months and these are fine but the samples fixed for 2-4 years do not uptake the Alcian Blue. I have taken it up to 1 hr in the 1% Alcian Blue solution followed by 10 min in 56*C Alkaline Alcohol solution. I do a 1hr Bouin's 50*C to start (I have also tried Bouin's at 60*C). Any ideas on how I can get the Alcian Blue to stain these excessively fixed samples or possibly a substitute? I need to see the proteoglycans/GAGs. Thank you Krista ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet